The reduction of Cr(VI) on the skin of living rats was investigated using the technique of in vivo electron paramagnetic resonance (EPR) spectroscopy with a surface coil resonator. Topical application of aqueous solution of Cr(VI) on the skin of rats generated the transient species, Cr(V). Partial removal of the stratum corneum increased the rates of formation and decay of Cr(V), as well as Cr(V) signal intensity. The results demonstrate that skin represents one of the routes for chromium to enter into animals and humans. The generation of reactive Cr(V) species in the reduction of Cr(VI) by the skin may play an important role in the mechanism of Cr(VI)-induced skin cancer. These findings indicate the potential for in vivo EPR spectroscopy to study the metabolism of paramagnetic reactive species in chemical and biochemical reactions occurring in/on the skin of both small and large animals, and possibly humans.